Exosomal miR-21-5p derived from endometrial stromal cells promotes angiogenesis by targeting TIMP3 in ovarian endometrial cysts

Endometriosis is a multifactorial gynecological disease, with angiogenesis as a key hallmark. The role of exosomal microRNAs (miRNAs) in endometriosis is not well understood. This study investigates differentially expressed exosomal miRNAs linked to angiogenesis in endometriosis, clarifies their molecular mechanisms, and identifies potential targets. Primary endometrial stromal cells (ESCs) were cultured, and exosomes were extracted. In a co-culture system, ESC-derived exosomes were taken up by human umbilical vein endothelial cells (HUVECs). Endometriosis implant-ESC-derived exosomes (EI-EXOs) significantly promoted HUVEC proliferation, migration and tube formation compared to normal endometrium-exosomes (NE-EXOs), a finding consistent in vivo in mice. MiRNA sequencing and bioinformatics identified differentially expressed miR-21-5p from EI-EXOs, confirmed by RT-qPCR. The miR-21-5p inhibitor or GW4869 attenuated EI-EXO-induced HUVEC proliferation, migration, and tube formation. TIMP3 overexpression diminished the pro-angiogenic effect of EI-EXOs, which was reversed by adding EI-EXOs or upregulating miR-21-5p. These findings validate the crosstalk between ESCs and HUVECs mediated by exosomal miR-21-5p, and confirm the miR-21-5p-TIMP3 axis in promoting angiogenesis in endometriosis. Key messages - ESC-derived exosomes were found to be taken up by recipient cells, i.e. HUVECs. Functionally, endometriosis implant-ESC-derived exosomes (EI-EXOs) could significantly promote the proliferation, migration and tube formation of HUVECs compared to normal endometrium-exosomes (NE-EXOs). - Through miRNA sequencing and bioinformatics analysis, differentially expressed miR-21-5p released by EI-EXOs was chosen, as confirmed by qRT-PCR. - miR-21-5p inhibitor or GW4869 was found to attenuate the proliferation, migration, and tube formation of HUVECs induced by EI-EXOs. In turn, TIMP3 overexpression diminished the pro-angiogenic effect of EI-EXOs, and this angiogenic phenotype was reversed once EI-EXOs were added or miR-21-5p was upregulated. Similar content being viewed by others Data availability Data sharing is not applicable to this article as no datasets were generated or analysed during the current study.

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Am J Obstet Gynecol 223(4):557 .e1-557.e11 Funding This work was supported by the National Natural Science Foundation of China (Grant No. 81971359), National Natural Science Foundation of Heilongjiang Province (Grant No. LH2019H027), and Key R&D project of Heilongjiang Province (Grant No. GA21C008) to G.Z., and National Natural Science Foundation of China (Grant No. 82101725) to Y.C. Author information Authors and Affiliations Contributions L.S. and G.Z. designed this study. L.S., Y.C., J.W., D.W., L.Y., X.W., Y.L., and J.G. conducted the experiments, analysed the data, and performed statistical analysis. L.S. wrote the manuscript with help from Y. C., J. W., G. Z. L. Y., X. W., and Y. L. provided scientific support and discussion. G.Z. critically reviewed the manuscript and provided scientific support for experiments. All authors revised and commented on the manuscript drafts. Corresponding author Ethics declarations Ethics statement The studies involving human specimens were reviewed and approved by the Ethical Committee of the First Affiliated Hospital of Harbin Medical University, and written informed consent was obtained from all participants. The animal study protocol was reviewed and approved by the Committee on the Ethical Use of Animals of the Harbin Medical University. Conflict of interest All the authors declare no competing interests. Additional information Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Supplementary Information Below is the link to the electronic supplementary material. Rights and permissions Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. About this article Cite this article Sun, L., Cheng, Y., Wang, J. et al. Exosomal miR-21-5p derived from endometrial stromal cells promotes angiogenesis by targeting TIMP3 in ovarian endometrial cysts. J Mol Med 102, 1327–1342 (2024). https://doi.org/10.1007/s00109-024-02483-z Received: Revised: Accepted: Published: Version of record: Issue date: DOI: https://doi.org/10.1007/s00109-024-02483-z